Part:BBa_K1604030

pncB

pncB encodes for the enzyme NAPRTase (nicotinic acid phosphoribosyl-transferase). It catalyzes the formation of nicotinate mono-nucleotide, a direct precursor of NAD, from NA.[1] [2] [3].

Usage and Biology

FIGURE 1. Biochemical pathway of NAD synthesis. PncB gene encodes the transcription of the NARPTase which catalyzes the formation of nicotinate mono-nucleotide from nicotinic acid. This is considered the limiting step of the synthesis of NAD.

FIGURE 2. PncB increases NAD levels by 126% (2.5 fold) and NADH levels by 44% (1.4 fold) when expressed in NEB10β. NAD and NADH levels were calculated with a colorimetric assay using the Sigma NAD /NADH quantification kit (MAK037) following the instructions described in the technical bulletin. The kit provides the measures of NAD levels indirectly from total levels of NAD + NADH and NADH only. Panel A. Colorimetric assay for NAD/NADH quantification. Panel B. NAD/NADH levels for three biological samples expressin BBa_K1604031 and one negative control expressing BBa_K731201.

For more information about the pncB (BBa_K1604030) characterization go on BBa_K1604031 page, or on Unitn iGEM 2015 project checking out our Wiki: Unitn iGEM wiki 2015

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1. Susana J. Berrıos-Rivera, K.-Y. San, and G. N. Bennett. (2002) Metabolic Engineering 4, 238–247

2. Marcel G. Wubbolts, Peter Terpstra, Jan B. van Beilen, Jaap Kingma, H. A. Rene Meesters, and Bernard Witholt. (1990) JBC, 265 (29), 17665-72

3. Ka-Yiu San, George N. Bennett, Susana J. Berríos-Rivera, Ravi V. Vadali, Yea-Tyng Yang, Emily Horton, Fred B. Rudolph, Berna Sariyar, and Kimathi Blackwood (2002) Metabolic Engineering, 4, 182–192

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1106
  
• 1000
  COMPATIBLE WITH RFC[1000]